Imactiv-3D

Introduction :

To fully exploit the capabilities of 3D in vitro or ex vivo models, it is essential to preserve the 3D regionalization level of analysis. In addition to confocal and two-photon, we developed a specific expertise in light sheet fluorescence microscopy (LSFM), a fast 3D imaging technique. Once the biological sample prepared, a sheet of laser light passes through it illuminating only a thin slice. A camera placed perpendicularly to the light sheet picks up the fluorescence signal, resulting in images of the illuminated region. This method has many advantages: in addition to being able to image a large range of fixed, live or cleared samples at a great resolution and depth, it also reduces acquisition duration and photodamage.

Service description :

• Sample handling, including, if necessary, labeling, conditioning for imaging, clearing.
• Image acquisition by our imaging experts, if necessary, multi-channel, with mosaicking, at different scales.
• Delivery of the images in different possible forms: stacks of images, video. Delivered through an online, secure, user-friendly storage solution. Imaging stored and sent using hard drives remains possible.

Imactiv-3D can process complex 2D-3D images with state-of-the-art and custom-made algorithms for restoration, segmentation, classification, quantification and visualization.

From sample conditioning and imaging to software development, each step can be tuned and adjusted on demand to best meet your requirements. 

Custom-based image processing algorithms are designed to perform specific jobs, including image restoration, segmentation and classification, based on approaches ranging from statistical modelling to deep learning. 

This framework enables to:

- reduce noise and blur
- improve resolution
- identify specific structures in complex environment
- track down subsets of samples in a supervised or unsupervised manner.

Quantitative results, together with 3D innovative media for visualization or marketing applications, are delivered through an online, secure, user-friendly storage solution. Imaging and quantification results stored and sent using hard drives remains possible.

Imactiv-3D brings expertise together to optimize the processing of your biological samples in order to achieve outstanding quality imaging.

For wide-field microscopy:
- Histological sections, cryosections
- Large range of labelling (dyes, antibodies, coloration)

For 3D imaging:
- Sample clearing and inclusion:
- In vitro 3D models (spheroids, organoids, bioprinting, RHE…)
- Ex vivo samples (biopsy, tissue, whole organ)
- Immuno-labelling in toto

Because of the ethical and societal considerations as well as the very high cost of animal testing, in vitro 3-D models that recapitulate normal or pathological human tissues are essential for understanding the intimate mechanisms of growth and for the development of new treatments. Spheroids or micro-tissues faithfully reproduce the organization of a tissue, recapitulating the cell-cell connections, cell-microenvironment and cellular heterogeneity found in the micro-regions of a tumor or organ. They are considered attractive models to assess the anti-proliferative activity of new drugs or genotoxic properties. The activity we offer is based on our strong expertise in the development of multicellular 3-D models from existing lines or from cells supplied by customers. We can support the assessment of a compound from 3-D cell culture, through the processing of 3-D models up to their three-dimensional exploration. We also propose culture in hypoxia and physoxia conditions of 3-D models grown either in suspension or in your scaffold.

The in vitro evaluation of cell viability and cell proliferation is a key step in the assessment of the biological activity of a new pharmacological compound or a natural product.

To reach this objective, Imactiv-3D offers its expertise in cell culture, cancer cell biology and pharmacology to test your compounds using 2D and 3D cell culture systems. 

We can provide two different approaches:

o   Classical molecular probes (WST1/Alamar blue…) can be combined with a fluorescent plate reader analysis to estimate EC50 or IC50 of your molecule/compound.

o   A PI labeling (or other specific labeling of cell death) combined with an automated imaging and cell counting analysis. This can be done together with other specific labeling such as proliferation.

It’s no secret that the success of any experiments depends on the quality of the starting material, and this is especially true for imaging biological samples. Sample preparation is crucial to obtain high quality, reproducible and reliable images. That’s why our team has developed innovative protocols to prepare biological samples by labelling and clearing for 3D and wide-field imaging. As every sample is different in terms of size, shape, state and physical properties, we work out the best conditions required for highest quality imaging.

- Quantitative image analysis driven by your need (effect of a compound for instance...), with development of specific numerical or qualitative criteria.

- Innovative and state-of-the-art quantification methods best designed for your study.

We can work with any kind of imaging modalities 

- All outputs available to present your results in the best possible ways: graphs, images, films...

- Various models for 3D rendering: surface, volume, mixed

- Original and persuasive means to visualize your data

See what we can do here 

Detect and visualize specific marker or cellular structure while preserving 3D sample morphology:

In toto sample labeling and handling for microscopy:

- Tunable in-house protocol

- Consecutive steps of permeabilization, saturation antibody incubation best suited for 3D samples

- Possibility of complementary dye staining if required

- Clearing

Our team can also take in charge the image acquisition in 3D if required (light sheet, confocal or two-photon microscopy) and the image analysis 

Imactiv-3D brings together experts in cell biology and imaging as well as experts in mathematics and image processing.
We propose you the association of these expertises for :
* Choose the best imaging modality to meet your needs
* Take charge of the imaging of your samples
* Advise you in your internal imaging procedures

Structured light microscopy is very useful for developing automated plate reading tests. Less resolute than confocal microscopy, it allows to meet low-cost analysis needs on thin samples (<200µm): 3D ultra-flat microscopy.
We have developed a strong expertise in the development of automated tests including :
* Thin cell culture (cell cluster, large cells such as adipocytes, scaffold culture)
* The application of treatment conditions
* Fluorescent markings
* Clearing compatible in multi-well plates if required
* Automated imaging with mosaicking and stitching to increase the analysis area, and image stacking to take into account the thickness of the samples.
Image processing for quality control and quantification (viability, proliferation, staticity etc.)

Confocal microscopy is particularly suitable for 3D exploration with excellent resolution of thin samples (<200µm).
We recommend this technology for the exploration and quantification of small biological structures (<5µm).
The systems at our disposal allow us to work in 96-well plates with automated protocols.
We have developed a strong expertise in the development of automated tests including :
* Thin cell culture (cell cluster, large cells such as adipocytes, scaffold culture)
* The application of treatment conditions
* Fluorescent markings
* Clearing compatible in multi-well plates if required
* Automated imaging with mosaicking and stitching to increase the analysis area, and image stacking to take into account the thickness of the samples.
Image processing for quality control and quantification (viability, proliferation, staticity etc.)